Aquaculture America 2020

February 9 - 12, 2020

Honolulu, Hawaii

DNA VACCINE TARGETING TO MATURE MYOSTATIN INCREASE GROWTH PERFORMANCES IN ORANGE-SPOTTED GROUPER Epinephelus coioides

Huirong Yang1, Jun Xia2, Yan Yang1, Qing Wang1, Huihong Zhao1, Jinzeng Yang3*
1 College of Marine Sciences, South China Agricultural University, Guangzhou 510640, China 2Institute of Veterinary Medicine, Xinjiang Academy of Animal Sciences, Urumqi 830000, China; 3Department of Human Nutrition, Food and Animal Sciences, University of Hawaii at Manoa, Honolulu, HI 96822, USA.

Myostatin (MSTN), also known as the growth differentiation factor-8 (GDF-8) is a member of the TGF-β superfamily, negatively regulates skeletal muscle growth. Depression of myostatin function leads to dramatic growth of muscle mass in mice, human and several livestock animals. The role of myostatin function in fish skeletal muscle growth was not consistent as there are several MSTN orthologs and its expression is not restricted to muscle. We isolated and identified myostatin DNA sequence from orange-spotted grouper Epinephelus coioides by using RACE technique. The myostatin cDNA of E. coioides consisted of 3782 nucleotides with a 99 bp 5'UTR, 1378 bp 3'UTR and an open reading frame (ORF) of 1131 bp, encoding for a protein of 376 amino acid residues. Myostatin mRNA are widely expressed in various tissues with the highest expression in the skeletal muscle. In an attempt to down-regulate myostatin, we constructed a DNA vaccine against myostatin (named PCS-MSTN) by cloning the hepatitis B surface antigen (HBsAg) S gene to E. coioides mature myostatin cDNA sequence in pcDNA3.1 vector. The recombinant vector PCS-MSTN was used to immunize E. coioides with body length 10cm and weight at approximate 25g over 3 times with interval of 2 weeks of vaccination. After 1 months of final vaccination, sera from vaccinated grouper showed the presence of specific antibodies against myostatin. In addition, the body weight of fish treated with 200 ng DNA vector was increased significantly. Myogenin mRNA expressions in skeletal muscle and brain tissue were increased in comparison with the controls. These preliminary results provide initial evidences that depression of myostatin function via DNA vaccine may promote E.coioides grouper growth performances.

Figure 1. Schematic diagram of the recombinant vector PCS-MSTN. DNA vaccine against myostatin (named PCS-MSTN) by cloning the hepatitis B surface antigen (HBsAg) S gene to Epinephelus coioides mature myostatin cDNA sequence in pcDNA3.1 vector.

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